The residue 179 is involved in product specificity of the Bacillus circulans DF 9R cyclodextrin glycosyltransferase

Applied Microbiology and Biotechnology
Hernán CostaSusana Ferrarotti

Abstract

Cyclodextrin glycosyltransferases (CGTases) are important enzymes in biotechnology because of their ability to produce cyclodextrin (CD) mixtures from starch whose relative composition depends on enzyme source. A multiple alignment of 46 CGTases and Shannon entropy analysis allowed us to find differences and similarities that could be related to product specificity. Interestingly, position 179 has Gly in all the CGTases except in that from Bacillus circulans DF 9R which possesses Gln. The absence of a side chain at that position has been considered as a strong requirement for substrate binding and cyclization process. Therefore, we constructed two mutants of this enzyme, Q179L and Q179G. The activity and kinetic parameters of Q179G remained unchanged while the Q179L mutant showed a different CDs ratio, a lower catalytic efficiency, and a decreased ability to convert starch into CDs. We show that position 179 is involved in CGTase product specificity and must be occupied by Gly--without a side chain--or by amino acid residues able to interact with the substrate through hydrogen bonds in a way that the cyclization process occurs efficiently. These findings are also explained on the basis of a structural model.

References

Jul 1, 1996·Biophysical Journal·B J Strait, T G Dewey
Oct 31, 1998·Protein Engineering·I N Shindyalov, P E Bourne
Jan 11, 2001·Biochimica Et Biophysica Acta· van der Veen BAL Dijkhuizen
May 30, 2002·Protein Engineering·Ivan Y TorshinRobert W Harrison
Feb 11, 2003·Proteins·Shandar AhmadAkinori Sarai
Apr 30, 2003·Protein Science : a Publication of the Protein Society·Björn Wallner, Arne Elofsson
Jun 3, 2004·Genome Research·Gavin E CrooksSteven E Brenner
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Oct 3, 2006·Cellular and Molecular Life Sciences : CMLS·M Machovic, S Janecek
Dec 2, 2006·International Journal of Pharmaceutics·Thorsteinn Loftsson, Dominique Duchêne
Dec 19, 2006·Bioresource Technology·Natalia SzermanSusana Alicia Ferrarotti
Apr 23, 2008·Current Protocols in Bioinformatics·Narayanan EswarAndrej Sali
Oct 8, 2008·Nucleic Acids Research·Brandi L CantarelBernard Henrissat
Oct 17, 2008·Carbohydrate Research·Hernán CostaMirtha Biscoglio de Jiménez Bonino
Apr 16, 2009·Applied Microbiology and Biotechnology·Ronan M KellyHans Leemhuis
Sep 19, 2009·Applied Microbiology and Biotechnology·Hans LeemhuisLubbert Dijkhuizen

❮ Previous
Next ❯

Citations

Jul 4, 2012·International Journal of Molecular Sciences·Poh Hong GohKian Mau Goh
Mar 3, 2017·Journal of Agricultural and Food Chemistry·Ye-Seul KooJae-Hoon Shim
Oct 10, 2019·Anais Da Academia Brasileira De Ciências·Soledad Caminata LandrielHernán Costa
Oct 9, 2013·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Yang YueShijun Qian

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.